Device capable of being used for preparing multi-layer cell sheet
By designing a fixture and a culture box fixing device, the problem of unstable cell sheet position in the preparation of multilayer cell sheets was solved, enabling the preparation of multilayer cell sheets without exogenous materials and simplifying the cleaning process, thereby improving preparation efficiency and safety.
Patent Information
- Application Number
- CN202423178590.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-23
- Publication Date
- 2026-02-03
- Estimated Expiration
- 2034-12-23
AI Technical Summary
In existing technologies, the cell sheet position cannot be fixed during the preparation of multilayer cell sheets, leading to the failure of multilayer cell sheet preparation. Furthermore, the use of thermosensitive polymer materials and gel extraction processes raises concerns about unknown cytotoxicity and safety.
A device comprising a clamp and a culture box was designed. The clamp consists of a clamp cover and a clamp base. Cell sheets are fixed by a limiting structure and a perforated opening to ensure that they do not move during the culture process. The design of the culture box and the clamp facilitates the fixation and cleaning of the cell sheets.
This technology enables the one-time preparation of multilayer cell sheets without relying on exogenous extracellular matrix, avoiding cell damage and cleaning complexity, and improving the cleaning efficiency and ease of operation of cell sheets.
Smart Images
Figure CN223866681U_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biotechnology, specifically relating to a device for preparing multilayer cell sheets. Background Technology
[0002] Cell sheets can be formed by continuously culturing cells to a high density or by inducing cell culture. Besides research in life sciences and clinical medicine, cell sheets can also be applied in clinical treatment. Although culture methods can create multi-layered cell sheets from a single layer, the growth of cultured cell sheets is limited by factors such as nutrients, metabolic products, and dissolved oxygen levels in the culture medium, making it difficult to grow cell sheets suitable for complex surgical procedures, such as those requiring sutures.
[0003] Furthermore, producing thick cell sheets requires culturing cell sheets on thermosensitive polymeric materials, then removing the cell sheets with a gel and stacking them, utilizing the gel's properties to bond the cell sheets. However, thermosensitive polymeric materials, such as poly(N-isopropylacrylamide), are cytotoxic and can also affect cell morphology. On the other hand, gel residues are easily left on the cell sheets during the gel removal process, and their safety is unknown. These challenges limit the clinical application of multilayer cell sheets, especially.
[0004] Therefore, it is necessary to provide a manufacturing technique for preparing multilayered cell sheets. However, cell sheets tend to move during the multilayering process, making it difficult for them to form effective connections. Utility Model Content
[0005] The purpose of this invention is to provide an apparatus for preparing multilayer cell sheets in order to solve at least one of the above-mentioned problems. This apparatus solves the problem of failure in preparing multilayer cell sheets when the cell sheet position cannot be fixed in the prior art without exogenous materials. It enables the preparation of multilayer cell sheets in one go without relying on exogenous extracellular matrix.
[0006] The objective of this utility model is achieved through the following technical solution:
[0007] The technical solution of this utility model is to propose an apparatus for preparing multilayer cell sheets, including a clamp and a culture box for placing the clamp;
[0008] The fixture includes a fixture cover plate and a fixture base, with the fixture cover plate connected to the upper part of the fixture base;
[0009] The fixture cover plate has multiple cover plate cutouts, and the fixture base has multiple base cutouts, with the cover plate cutouts and base cutouts having the same shape at the same location;
[0010] The clamp base is equipped with limiting structures on all four sides. The cell slices are pressed between the limiting structures by the clamp cover plate and thus fixed between the clamp cover plate and the clamp base.
[0011] Furthermore, the cutouts in the cover plate and the base plate at the same location have the same shape, and these cutouts are used to allow the culture medium in the culture box to pass through the clamp.
[0012] Furthermore, the hinged edge of the fixture cover plate is hinged to the hinged edge of the fixture base.
[0013] Furthermore, one end of each of the two adjacent sides of the hinged edge of the clamp cover plate extends outward as an extended end, a pivot is provided on the outer side of the extended end, and a cover plate locking position is provided on the outer side of the other end, with the pivot and the cover plate locking position protruding outward;
[0014] The two sides of the two adjacent sides of the hinged edge of the clamp base are provided with inwardly recessed cover plate slots. The cover plate slots are rotatably connected to the rotating shaft and are engaged with the cover plate slots.
[0015] Furthermore, the limiting structure of the fixture base includes a limiting strip and a limiting plate.
[0016] A limiting strip is provided on the edge of the fixture base, and multiple protrusions are provided on the surface of the limiting strip;
[0017] The limiting plate is a sidewall formed by extending upward from the side of the fixture base, and the distance between the relative limiting plates formed by the two opposite sides of the fixture base is greater than the distance between the opposite sides of the fixture cover plate.
[0018] Furthermore, the limiting strip is used to restrict the movement of the cell sheet in the clamp, and the protrusions of the limiting strip can be wavy, continuous, or discontinuous.
[0019] Furthermore, one side of the clamp cover extends outward to form a clamping plate, and no limiting plate is provided at the connection between the clamp base and the clamping plate.
[0020] Furthermore, the fixture base is equipped with multiple support bars, and the surface of the support bars is raised.
[0021] The limiting plate, support strip, and limiting strip work together to restrict the movement of the cell sheet in the fixture.
[0022] Furthermore, the culture box has multiple clamping and limiting components on the inner side of its opening and multiple outwardly protruding culture box slots on the outer side. The clamping and limiting components are used to fix the clamps and prevent them from shaking inside the culture box.
[0023] Furthermore, the apparatus for preparing multilayer cell sheets also includes a culture box lid, which is snapped onto the outside of the culture box opening.
[0024] Furthermore, multiple lid slots are provided on the inner side of the culture box lid, and the culture box lid and the culture box are engaged with each other through the culture box locking position and the lid slots.
[0025] Furthermore, the culture box slots correspond one-to-one with the box lid slots.
[0026] Furthermore, the number of the culture box slots and the box cover slots is 4-8.
[0027] Furthermore, the clamp is placed with its hinged edge or two adjacent sides facing downwards in the culture box.
[0028] Furthermore, the clamp is fixed or suspended in the culture box.
[0029] Furthermore, a clamp base limiter is provided at the bottom of the culture box, and the clamp base limiter is connected to the clamp.
[0030] Furthermore, the surface of the clamp base limiter that contacts the clamp is provided with multiple through holes, creating a gap between the bottom of the clamp base limiter and the clamp, and also a gap between the bottom of the clamp base limiter and the bottom of the inner side of the culture box. The through holes and gaps allow for free osmotic exchange between the culture medium in the clamp and the culture medium in the culture box, maintaining the freshness of the culture medium around the cell sheets within the clamp, and also facilitating medium replacement operations.
[0031] Furthermore, the device is made of medical-grade radiation-resistant plastic, including any one of PETG, PVDF, PES, PEEK, and PSU materials.
[0032] Compared with the prior art, the present invention has the following advantages:
[0033] 1. The device of this utility model is equipped with a clamp and culture box that can simultaneously culture multiple cell sheets. The cell sheets can be fixed in the position of the clamp by the cell sheet fixing component. During the culture process, the cell sheets are kept in contact, thereby generating intercellular connections or secreting extracellular matrix. Multilayer cell sheets can be prepared in one go without relying on exogenous extracellular matrix.
[0034] 2. This utility model fixes the cell sheet in the clamp, and after the cell culture is completed, the clamp can be directly picked up and placed in physiological saline for quick cleaning. This avoids damage to the cell sheet caused by directly clamping it, improves the cleaning efficiency of the cell sheet, simplifies the cleaning work, and the hollow opening of the clamp is conducive to placing the clamp together with the cell sheet in other working solutions, which simplifies the operation steps.
[0035] 3. This utility model, by providing a clamp cover and a clamp base for the clamp, makes it easy to distinguish the front and back of the clamp on which the cell slices are placed. The front and back of the cell slices can be directly distinguished through the clamp cover and the base, eliminating the need to mark the cell slices, reducing the complexity of the production process. After cleaning, the top cover can be opened directly without distinguishing the front and back, which facilitates the application and operation of the cell slices and greatly improves efficiency. Attached Figure Description
[0036] Figure 1 This is a schematic diagram of the fixture of the present invention.
[0037] Figure 2 This is a schematic diagram of the structure of the clamp cover plate of the device of this utility model.
[0038] Figure 3 This is a schematic diagram of the fixture base of the present invention.
[0039] Figure 4 This is a schematic diagram of the structure of the culture box and clamp of the device of this utility model.
[0040] Figure 5 This is a schematic diagram of the overall structure of the culture box of the present invention.
[0041] Figure 6 This is a schematic diagram of the overall structure of the culture box in Embodiment 2 of the present invention.
[0042] Figure 7 This is a cross-sectional schematic diagram of the device in Embodiment 2 of this utility model.
[0043] In the diagram: 1-Clamp; 2-Clamp cover plate; 21-Cover plate slot; 22-Cover plate cutout; 23-Spindle; 24-Clamp plate; 3-Clamp base; 31-Cover plate slot; 32-Limiting strip; 33-Supporting strip; 34-Base cutout; 35-Limiting plate; 4-Cultivation box; 41-Clamp limiting component; 42-Cultivation box slot; 5-Cultivation box cover; 6-Clamp base limiter. Detailed Implementation
[0044] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments. This embodiment is based on the technical solution of the present invention and provides detailed implementation methods and specific operating procedures; however, the scope of protection of the present invention is not limited to the following embodiments.
[0045] In the description of this utility model, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer," etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the accompanying drawings and are only for the convenience of describing this utility model and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of this utility model. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.
[0046] In the description of this utility model, it should be noted that, unless otherwise explicitly specified and limited, the terms "installation," "connection," and "joining" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of two components. Those skilled in the art can understand the specific meaning of the above terms in this utility model based on the specific circumstances.
[0047] Unless otherwise specified, the functional components or structures in the following embodiments or examples are conventional components or structures used in the art to achieve the corresponding functions.
[0048] Example 1
[0049] like Figures 1-5 As shown, an apparatus for preparing multilayer cell sheets, made of PETG material, includes a clamp 1, a culture box 4 for placing the clamp 1, and a culture box cover 5.
[0050] like Figure 1 As shown, the fixture 1 includes a fixture cover plate 2 and a fixture base 3. The fixture cover plate 2 is connected to the upper part of the fixture base 3, and the hinge side of the fixture cover plate 2 is hinged to the hinge side of the fixture base 3.
[0051] like Figure 2 As shown, the clamp cover plate 2 is provided with multiple cover plate cutouts 22; one end of each of the two adjacent sides of the hinged edge of the clamp cover plate 2 extends outward as an extension end, and an outwardly protruding pivot 23 is provided on the outer side of the extension end; an outwardly protruding cover plate slot 21 is provided on the outer side of the other end; a clamping plate 24 extends outward from one side of the clamp cover plate 2.
[0052] like Figure 3As shown, the clamp base 3 has inwardly recessed cover plate slots 31 on the outer sides of the two adjacent sides of the hinge edge; the clamp base 3 has limiting structures on all four sides, including limiting strips 32 and limiting plates 35. The limiting strips 32 are located at the edge of the clamp base 3, and the surface of the limiting strips 32 has multiple wavy protrusions; the limiting plates 35 are sidewalls formed by the upward extension of the side of the clamp base 3, and the distance between the relative limiting plates 35 formed by the two opposite sides of the clamp base 3 is greater than the distance between the opposite sides of the clamp cover plate 2; the clamp base 3 has multiple support strips 33, and the surface of the support strips 33 is raised; the limiting plates 35, support strips 33 and limiting strips 32 are used together to restrict the movement of cell sheets in the clamp 1; the clamp base 3 has multiple base cutouts 34;
[0053] The cover plate slot 31 is rotatably connected to the rotating shaft 23, and the cover plate slot 31 is engaged with the cover plate slot 21; the cover plate cutout 22 and the base cutout 34 at the same position have the same shape; no limiting plate 35 is provided at the connection between the clamp base 3 and the clamp plate 24; the cell slices are pressed between the limiting structures by the clamp cover plate 2, and thus fixed between the clamp cover plate 2 and the clamp base 3;
[0054] like Figure 4 As shown, the culture box 4 has multiple clamping and limiting parts 41 on the inner side of the box opening and multiple outwardly protruding culture box slots 42 on the outer side.
[0055] Multiple lid slots are provided on the inner side of the culture box lid 5. The culture box lid 5 and the culture box 4 are engaged with the lid slots through the culture box locking position 42.
[0056] like Figure 5 As shown, this embodiment provides an apparatus for preparing multilayer cell sheets, with clamp 1 measuring 37.5x37.2x6.6mm and culture box 4 measuring 43.2x14.3x39.4mm, suitable for culturing 30x30mm cell sheets.
[0057] like Figure 4 As shown, when using, stack the cell sheets and place them between the limiting strip 32 and the limiting plate 35 in clamp 1. Cover with clamp cover 2, place clamp 1 into culture box 4, and fix clamp 1 with clamp limiting piece 41. Pour 35mL of culture medium into culture box 4, cover with culture box cover 5, and place culture box 4 in a 37℃, 5.0% CO2 incubator. Change the medium every other day and culture for 14 days until the cell sheets connect into single cell sheets. After culture, remove culture box cover 5, use sterile forceps to pick up clamp 1, and quickly shake and wash it three times in 500ml sterile physiological saline. After washing, gently open clamp cover and it is ready for use.
[0058] Example 2
[0059] like Figures 1-3 , Figure 6 as well as Figure 7 As shown, an apparatus for preparing multilayer cell sheets is made of PVDF material and includes a clamp 1, a culture box 4 for placing the clamp 1, and a culture box cover 5.
[0060] like Figure 1 As shown, the fixture 1 includes a fixture cover plate 2 and a fixture base 3. The fixture cover plate 2 is connected to the upper part of the fixture base 3, and the hinge side of the fixture cover plate 2 is hinged to the hinge side of the fixture base 3.
[0061] like Figure 2 As shown, the clamp cover plate 2 is provided with multiple cover plate cutouts 22; one end of each of the two adjacent sides of the hinged edge of the clamp cover plate 2 extends outward as an extension end, and an outwardly protruding pivot 23 is provided on the outer side of the extension end; an outwardly protruding cover plate slot 21 is provided on the outer side of the other end; a clamping plate 24 extends outward from one side of the clamp cover plate 2.
[0062] like Figure 3 As shown, the clamp base 3 has inwardly recessed cover plate slots 31 on the outer sides of the two adjacent sides of the hinge edge; the clamp base 3 has limiting structures on all four sides, including limiting strips 32 and limiting plates 35. The limiting strips 32 are located at the edge of the clamp base 3, and the surface of the limiting strips 32 has multiple wavy protrusions; the limiting plates 35 are sidewalls formed by the upward extension of the side of the clamp base 3, and the distance between the relative limiting plates 35 formed by the two opposite sides of the clamp base 3 is greater than the distance between the opposite sides of the clamp cover plate 2; the clamp base 3 has multiple support strips 33, and the surface of the support strips 33 is raised; the limiting plates 35, support strips 33 and limiting strips 32 are used together to restrict the movement of cell sheets in the clamp 1; the clamp base 3 has multiple base cutouts 34;
[0063] The cover plate slot 31 is rotatably connected to the rotating shaft 23, and the cover plate slot 31 is engaged with the cover plate slot 21; the cover plate cutout 22 and the base cutout 34 at the same position have the same shape; no limiting plate 35 is provided at the connection between the clamp base 3 and the clamp plate 24; the cell slices are pressed between the limiting structures by the clamp cover plate 2, and thus fixed between the clamp cover plate 2 and the clamp base 3;
[0064] The culture box 4 has multiple outwardly protruding culture box slots on the outer side of its opening; a clamp base limiter 6 is provided at the bottom of the culture box 4, and the clamp base limiter 6 is connected to the clamp 1; the culture box cover 5 has multiple cover slots on its inner side, such as... Figure 6 As shown, the culture box lid 5 and the culture box 4 are engaged with the lid slot via the culture box locking mechanism.
[0065] Figure 7 This is a cross-sectional schematic diagram of the device in Example 2, as shown below. Figure 7As shown, this embodiment provides an apparatus for preparing multilayer cell sheets. The clamp 1 is placed at the bottom of the culture box 4. The size of the clamp 1 is 37.5x37.2x6.6mm, and the size of the culture box 4 is 43.2x43.2x14.3mm, which is suitable for culturing 30x30mm cell sheets.
[0066] like Figure 7 As shown, during use, stack the cell sheets and place them between the limiting strip 32 and the limiting plate 35 in clamp 1. Cover with clamp cover 2 and fix clamp 1 with clamp cover 2 facing upwards in the clamp base limiting device 6 at the bottom of the culture box 4. Clamp base limiting device 6 is used to fix the clamp and prevent it from shifting during culture. There is a gap between the bottom of clamp base limiting device 6 and the bottom of the inner side of culture box 4, and a gap between the bottom of clamp base limiting device 6 and clamp 1, allowing the culture medium in clamp 1 to freely exchange with the culture medium in culture box 4, maintaining the freshness of the culture medium around the cell sheets in clamp 1, and also facilitating medium replacement. Pour 20mL of culture medium into culture box 4, cover with culture box lid 5, and place culture box 4 at 37℃, 5.0% In a CO2 incubator, change the medium every other day and culture for 14 days until the cell sheets connect into single cell sheets. After culture, remove the culture box cover 5, use sterile tweezers to pick up the clamp 1, and quickly shake and clean it three times in 500ml of sterile physiological saline. After cleaning, gently open the clamp cover 2 and it is ready for use.
[0067] The above description of the embodiments is provided to enable those skilled in the art to understand and use the utility model. It will be apparent to those skilled in the art that various modifications can be easily made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present utility model is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present utility model without departing from its scope should be within the protection scope of the present utility model.
Claims
1. An apparatus for preparing multilayer cell sheets, characterized in that, Includes a clamp (1) and a culture box (4) for placing the clamp (1); The clamp (1) includes a clamp cover plate (2) and a clamp base (3), with the clamp cover plate (2) connected to the upper part of the clamp base (3); The clamp cover plate (2) is provided with multiple cover plate cutouts (22), and the clamp base (3) is provided with multiple base cutouts (34); The clamp base (3) has limiting structures on all four sides. The cell slices are pressed between the limiting structures by the clamp cover plate (2) and thus fixed between the clamp cover plate (2) and the clamp base (3).
2. The apparatus for preparing multilayer cell sheets according to claim 1, characterized in that, The hinged edge of the clamp cover plate (2) is hinged to the hinged edge of the clamp base (3).
3. The apparatus for preparing multilayer cell sheets according to claim 2, characterized in that, The two adjacent sides of the clamp cover plate (2) extend outwards as the extension end. A pivot (23) is provided on the outside of the extension end, and a cover plate slot (21) is provided on the outside of the other end. The pivot (23) and the cover plate slot (21) protrude outwards. The clamp base (3) has inwardly recessed cover plate slots (31) on the outer sides of the two adjacent sides of the hinge edge. The cover plate slots (31) are rotatably connected to the rotating shaft (23) and are engaged with the cover plate slots (21).
4. The apparatus for preparing multilayer cell sheets according to claim 1, characterized in that, The limiting structure of the fixture base (3) includes a limiting strip (32) and a limiting plate (35). A limiting strip (32) is provided on the edge of the fixture base (3), and multiple protrusions are provided on the surface of the limiting strip (32); The limiting plate (35) is a side wall formed by extending upward from the side of the fixture base (3), and the distance between the relative limiting plates (35) formed by the two opposite sides of the fixture base (3) is greater than the distance between the opposite sides of the fixture cover plate (2).
5. The apparatus for preparing multilayer cell sheets according to claim 4, characterized in that, One side of the clamp cover plate (2) extends outward to form a clamp plate (24), and no limiting plate (35) is provided at the connection between the clamp base (3) and the clamp plate (24).
6. The apparatus for preparing multilayer cell sheets according to claim 1, characterized in that, The fixture base (3) is provided with multiple support bars (33), and the surface of the support bars (33) is raised.
7. The apparatus for preparing multilayer cell sheets according to claim 1, characterized in that, The culture box (4) has multiple clamping and limiting parts (41) on the inner side of the box opening and multiple outwardly protruding culture box slots (42) on the outer side.
8. The apparatus for preparing multilayer cell sheets according to claim 7, characterized in that, The apparatus for preparing multilayer cell sheets also includes a culture box cover (5), which is snapped onto the outside of the opening of the culture box (4).
9. The apparatus for preparing multilayer cell sheets according to claim 8, characterized in that, Multiple lid slots are provided on the inner side of the culture box lid (5). The culture box lid (5) and the culture box (4) are engaged with the lid slots through the culture box slots (42).
10. The apparatus for preparing multilayer cell sheets according to claim 1, characterized in that, The bottom of the culture box (4) is provided with a clamp base limiter (6), which is connected to the clamp (1).